A New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco

Owing to the economic value of its foliage, tobacco (Nicotiana tabacum) is cultivated all across the world. For the detection of genetically modified (GM) tobacco, there is a lack of universal standard material which ultimately limits the detection methods because the accuracy and comparability of t...

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Autores principales:	Jing Yu, Xiaolian Zhang, Muhammad Faheem Adil, Bo Lei, Mengao Jia, Huina Zhao, Shizhou Yu, Jiemin Liu, Yushuang Guo, Imran Haider Shamsi
Formato:	article
Lenguaje:	EN
Publicado:	Hindawi-Wiley 2021
Materias:	Nutrition. Foods and food supply TX341-641
Acceso en línea:	https://doaj.org/article/d61a5e220e9243d78b74a92f309d887d
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spelling	oai:doaj.org-article:d61a5e220e9243d78b74a92f309d887d2021-11-29T00:56:29ZA New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco1745-455710.1155/2021/3220013https://doaj.org/article/d61a5e220e9243d78b74a92f309d887d2021-01-01T00:00:00Zhttp://dx.doi.org/10.1155/2021/3220013https://doaj.org/toc/1745-4557Owing to the economic value of its foliage, tobacco (Nicotiana tabacum) is cultivated all across the world. For the detection of genetically modified (GM) tobacco, there is a lack of universal standard material which ultimately limits the detection methods because the accuracy and comparability of the results cannot be ensured. Here, we prepared a reference plasmid “pGMT27” for the detection of GM tobacco, which was 18,296 bp in length harboring two of the tobacco endogenous and seven exogenous genes. By using qualitative PCR test for the nine genes, 10 copies were used for plasmid sensitivity. In the quantitative real-time PCR (qPCR) assays with pGMT27 as a calibrator, the reaction efficiencies for P-35S and NR were 101.427% and 98.036%, respectively, whereas the limit of detection (LOD) and limit of quantification (LOQ) were 5 copies and 10 copies per reaction. For standard deviation (SD) and relative standard deviation (RSD) of the Ct values, the repeatability values were from 0.04 to 0.42 and from 0.18% to 1.29%, respectively; and the reproducibility values were from 0.04 to 0.39 and from 0.18% to 1.14%, respectively. For the unknown sample test, the average conversion factor (Cf) was 0.39, and the accuracy bias was from −15.55% to 1.93%; for precision, the SD values ranged from 0.02 to 0.62, while RSD values were from 1.34% to 10.6%. We concluded that using the pGMT27 plasmid as a calibrator provided a highly efficient transgenic detection method for flue-cured tobacco.Jing YuXiaolian ZhangMuhammad Faheem AdilBo LeiMengao JiaHuina ZhaoShizhou YuJiemin LiuYushuang GuoImran Haider ShamsiHindawi-WileyarticleNutrition. Foods and food supplyTX341-641ENJournal of Food Quality, Vol 2021 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Nutrition. Foods and food supply TX341-641
spellingShingle	Nutrition. Foods and food supply TX341-641 Jing Yu Xiaolian Zhang Muhammad Faheem Adil Bo Lei Mengao Jia Huina Zhao Shizhou Yu Jiemin Liu Yushuang Guo Imran Haider Shamsi A New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco
description	Owing to the economic value of its foliage, tobacco (Nicotiana tabacum) is cultivated all across the world. For the detection of genetically modified (GM) tobacco, there is a lack of universal standard material which ultimately limits the detection methods because the accuracy and comparability of the results cannot be ensured. Here, we prepared a reference plasmid “pGMT27” for the detection of GM tobacco, which was 18,296 bp in length harboring two of the tobacco endogenous and seven exogenous genes. By using qualitative PCR test for the nine genes, 10 copies were used for plasmid sensitivity. In the quantitative real-time PCR (qPCR) assays with pGMT27 as a calibrator, the reaction efficiencies for P-35S and NR were 101.427% and 98.036%, respectively, whereas the limit of detection (LOD) and limit of quantification (LOQ) were 5 copies and 10 copies per reaction. For standard deviation (SD) and relative standard deviation (RSD) of the Ct values, the repeatability values were from 0.04 to 0.42 and from 0.18% to 1.29%, respectively; and the reproducibility values were from 0.04 to 0.39 and from 0.18% to 1.14%, respectively. For the unknown sample test, the average conversion factor (Cf) was 0.39, and the accuracy bias was from −15.55% to 1.93%; for precision, the SD values ranged from 0.02 to 0.62, while RSD values were from 1.34% to 10.6%. We concluded that using the pGMT27 plasmid as a calibrator provided a highly efficient transgenic detection method for flue-cured tobacco.
format	article
author	Jing Yu Xiaolian Zhang Muhammad Faheem Adil Bo Lei Mengao Jia Huina Zhao Shizhou Yu Jiemin Liu Yushuang Guo Imran Haider Shamsi
author_facet	Jing Yu Xiaolian Zhang Muhammad Faheem Adil Bo Lei Mengao Jia Huina Zhao Shizhou Yu Jiemin Liu Yushuang Guo Imran Haider Shamsi
author_sort	Jing Yu
title	A New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco
title_short	A New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco
title_full	A New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco
title_fullStr	A New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco
title_full_unstemmed	A New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco
title_sort	new reference plasmid “pgmt27” provides an efficient transgenic detection method for flue-cured tobacco
publisher	Hindawi-Wiley
publishDate	2021
url	https://doaj.org/article/d61a5e220e9243d78b74a92f309d887d
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A New Reference Plasmid “pGMT27” Provides an Efficient Transgenic Detection Method for Flue-Cured Tobacco

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